Illumina NovaSeq 6000 paired end sequencing; Single-cell transcription profiling human dorsal forebrain organoids treated with Hyper-IL-6
Single-cell transcription profiling human dorsal forebrain organoids treated with Hyper-IL-6
Dorsal forebrain organoids were enzymatically dissociated using the Worthington Papain Dissociation System kit (Worthington Biochemical, Cat. no. LK003150). Multiplexing was performed by 3' CellPlex Kit Set A (10x Genomics, Cat. no. PN-1000261) following the manufacturer's instructions. Dead Cell Removal (Miltenyi, Cat. no. 130-090-101) was performed following the manufacturer's instructions. Samples were subjected to multiplexing using 3' CellPlex Kit Set A (10x Genomics, Cat. no. PN-1000261) following the manufacturer's instructions. For Vehicle samples, CMO301 (BIONi010-C) and CMO302 (HMGU1) were used, for Hyper-IL-6-treated samples, CMO303 (BIONi010-C) and CMO304 (HMGU1) were used. Finally, the cells were quantified and pooled in equal proportions with CMO301 and CMO302 denoted as sample 1 and CMO303 and CMO304 denoted as sample 2. Approximately 16,500 cells per channel were loaded onto a Chromium Single Cell 3′ Chip (10x Genomics, Cat. no. PN-120236) and processed through the Chromium controller to generate single-cell gel beads in emulsion. Library preparation was carried out according to the manufacturer's protocol. Feature barcode libraries were created using a 10X accessory kit.